Cytological and bacteriological sampling from filters used for embryo recovery to evaluate the uterine status of donor mares

While testing for uterine bacterial infection is usually performed prior to artificial insemination (AI), samples taken during or after embryo flushing are generally not assessed either in subfertile and old mares or in fertile mares, even though knowledge of the status of the uterine environment in which the embryo is to develop would help to predict the outcome of embryo transfer programmes. The presence of bacteria and inflammatory cells in the liquid retained in the filter after uterine flushing in donors was determined at the moment of embryo recovery. Primarily, a group of mares (n = 8) displaying evident clinical signs of endometritis was selected to evaluate the cytological and bacteriological findings in filters after uterine flushing and in uterine cotton swabs. Two uterine samples (for cytological and bacterial evaluation) were taken with cotton swabs and, subsequently, the uterus was flushed and the efflux was also subjected to bacteriological and cytological analysis. Later, a group of donors (n = 20) was also involved to evaluate the presence of bacteria and polymorphonuclear leukocytes (PMN). After embryo flushing and collection, the efflux retained in the filter was evaluated by cytology and bacteriology. A sterile cotton swab was then scrubbed on the filter mesh, and a bacterial culture was performed. The embryo recovery rate was 30% (n = 6); Escherichia coli was isolated in one efflux sample collected from embryo-productive flushings, while the other five samples were negative by culture. Bacterial growth (not considered as contamination) was observed in a total of three samples, although no inflammatory cells were detected. Bacteria were isolated in endometrial samples collected after embryo flushing in donor mares, although inflammatory cells were never present in the uterus of mares from which embryos were recovered. In the absence of clinical signs, cytological and/or bacteriological samplings are not very useful for estimating the success of embryo recovery in donor mares, but evaluation of the filter and efflux after uterine flushing in donors may provide valuable information regarding uterine status at embryo collection

Bayesian Analysis of the Effects of Olive Oil-Derived Antioxidants on Cryopreserved Buck Sperm Parameters

The present study evaluates the effect of olive oil-derived antioxidants, hydroxytyrosol (HT) and 3,4-dihydroxyphenylglycol (DHPG), on cryopreserved caprine sperm using Bayesian in- ference of ANOVA. For this proposal, sperm was collected, pooled and diluted in freezing media supplemented with different concentrations of HT, DHPG and the mixture (MIX) of both antioxidants. Sperm motility, viability, acrosome integrity, mitochondrial status, and lipid peroxidation (LPO) were assessed in fresh and frozen-thawed sperm samples. The results provided evidence that HT at low concentrations improves sperm motility and viability, and reduces the LPO. Contrastingly, DHPG and MIX exert a positive effect by reducing LPO values as concentration increases. Additionally, mi- tochondrial potential was reduced when samples were supplemented with HT at low concentrations and mixture of both antioxidants. Conclusively, the addition of olive oil-derived antioxidants (HT at 10 μg/mL and DHPG at 30 μg/mL) implements a protective effect in cryopreserved buck sperm. Bayesian analysis alternatives offer new possibilities to determine the repercussion of antioxidants on sperm, both quantitatively and qualitatively

Effect of three commercial extenders on sperm motility and fertility in liquid ram semen stored at 15 °C or 5 °C

The effect of different extenders on sperm motility and fertility was evaluated during liquid storage of ram semen at 5 °C and 15 °C. The semen was collected, pooled and diluted in three commercial extenders: Inra 96® (INRA) based on skimmed milk, Biladyl® A fraction (BIL) based on egg yolk, and Ovixcell® (OVIX) based on soybean lecithin. Then, sperm motility was evaluated at 0, 6, 24, 48, 72 and 96 h. In order to evaluate fertility, samples stored at 15 °C were used after dilution in INRA and OVIX. Results showed that progressive motility was significantly higher up to 72 h of storage in sperm samples maintained at 5 °C in comparison with 15 °C, similarly for each tested diluent. When samples were stored at 5 °C in OVIX, kinematic parameters such as velocity (except curvilinear velocity, VCL), trajectory [linearity (LIN), straightness (STR), wobble (WOB)], amplitude of lateral head displacement (ALH) and beat/cross frequency (BCF) were higher than in INRA and BIL. No significant differences in pregnancy rate were detected between INRA (62.6%) and OVIX (58.9%). In conclusion, liquid storage at 5 °C with OVIX extender is an interesting option since non-animal components are used, and this extender offers similar in vitro and in vivo efficacy as other extenders containing animal components

CIBERER : Spanish national network for research on rare diseases: A highly productive collaborative initiative

Altres ajuts: Instituto de Salud Carlos III (ISCIII); Ministerio de Ciencia e Innovación.CIBER (Center for Biomedical Network Research; Centro de Investigación Biomédica En Red) is a public national consortium created in 2006 under the umbrella of the Spanish National Institute of Health Carlos III (ISCIII). This innovative research structure comprises 11 different specific areas dedicated to the main public health priorities in the National Health System. CIBERER, the thematic area of CIBER focused on rare diseases (RDs) currently consists of 75 research groups belonging to universities, research centers, and hospitals of the entire country. CIBERER's mission is to be a center prioritizing and favoring collaboration and cooperation between biomedical and clinical research groups, with special emphasis on the aspects of genetic, molecular, biochemical, and cellular research of RDs. This research is the basis for providing new tools for the diagnosis and therapy of low-prevalence diseases, in line with the International Rare Diseases Research Consortium (IRDiRC) objectives, thus favoring translational research between the scientific environment of the laboratory and the clinical setting of health centers. In this article, we intend to review CIBERER's 15-year journey and summarize the main results obtained in terms of internationalization, scientific production, contributions toward the discovery of new therapies and novel genes associated to diseases, cooperation with patients' associations and many other topics related to RD research

CIBERER: Spanish national network for research on rare diseases: A highly productive collaborative initiative

13 páginas,1 figura, 3 tablas, 1 apéndice. Se extraen los autores pertenecientes a The CIBERER network que trabajan en Centros del CSIC del Appendix ACIBER (Center for Biomedical Network Research; Centro de Investigación Biomédica En Red) is a public national consortium created in 2006 under the umbrella of the Spanish National Institute of Health Carlos III (ISCIII). This innovative research structure comprises 11 different specific areas dedicated to the main public health priorities in the National Health System. CIBERER, the thematic area of CIBER focused on rare diseases (RDs) currently consists of 75 research groups belonging to universities, research centers, and hospitals of the entire country. CIBERER's mission is to be a center prioritizing and favoring collaboration and cooperation between biomedical and clinical research groups, with special emphasis on the aspects of genetic, molecular, biochemical, and cellular research of RDs. This research is the basis for providing new tools for the diagnosis and therapy of low-prevalence diseases, in line with the International Rare Diseases Research Consortium (IRDiRC) objectives, thus favoring translational research between the scientific environment of the laboratory and the clinical setting of health centers. In this article, we intend to review CIBERER's 15-year journey and summarize the main results obtained in terms of internationalization, scientific production, contributions toward the discovery of new therapies and novel genes associated to diseases, cooperation with patients' associations and many other topics related to RD research.This study has been funded by Instituto de Salud Carlos III (ISCIII) and Spanish Ministry of Science and InnovationPeer reviewe

Sperm subpopulations and quality in fractions obtained after single layer centrifugation in fresh normospermic ram samples

La técnica de centrifugación de una sola capa (SLC) ha sido desarrollada para seleccionar la mejor población de espermatozoides en un eyaculado con la finalidad de aumentar las tasas de fertilización mediante la inseminación artificial o fertilización in vitro. Se procesaron muestras normospérmicos de semen de carnero que contenían 800 y 3000 × 106 espermatozoides/ml (C800 y C3000, respectivamente) utilizando SLC. Se separaron tres fracciones de espermatozoides en cada muestra y se analizó el rendimiento, la calidad y las subpoblaciones de espermatozoides en cada una. En las muestras de C800, la tasa de recuperación de espermatozoides no varió entre las fracciones. Sin embargo, en las C3000, la fracción superior (F1) contenía más espermatozoides que la fracción inferior (F3). La F1 en las C3000 tenía un porcentaje mayor de espermatozoides (53.2 %) que, en las C800, mientras que en la F2 de las C3000 el porcentaje de espermatozoides recuperados fue menor (25.2 %) que en la F2 de las C800 (45.4 %). Aplicando los parámetros de motilidad de los espermatozoides se identificaron tres subpoblaciones de espermatozoides: SP1, de baja velocidad y sin movimiento progresivo (19.1 %); SP2, rápidos y progresivos (43.7 %); y SP3, rápidos, pero con movimiento no lineal (37.2 %). Si bien SLC se ha utilizado para la separación de espermatozoides en muestras de semen subóptimas y/o de baja concentración de espermatozoides, los resultados indican que SLC no es eficiente en la separación de diferentes poblaciones de espermatozoides en muestras normospérmicas de semen de carnero con altas concentraciones de espermatozoides.Single layer centrifugation (SLC) technique has been developed to select the best sperm population in the ejaculate in order to increase the fertilization rates by artificial insemination or in vitro fertilization. Normospermic ram semen samples containing 800 and 3,000 × 106 sperms/ml (C800 and C3000, respectively) were processed by SLC. Three sperm fractions were separated in each sample following silica-coloidal sperm centrifugation and sperm yield, quality and subpopulations were analyzed in each one. In C800 group, the sperm recovery rate did not vary in any studied fraction, but when samples were highly concentrated (C3000) the top fraction (F1) contained significantly higher spermatozoa than bottom fraction (F3). Also, it was observed that F1 in C3000 had got a significantly higher percentage of spermatozoa (53.2 %) than in C800, while the quantity of spermatozoa recovered in fraction 2 was lower (25.2 % vs 45.4 %). Based on the sperm motility parameters, three sperm subpopulations were identified: SP1, low velocity spermatozoa showing no progressive movement (19.1 %); SP2, rapid and progressive spermatozoa (43.7 %); and SP3, rapid spermatozoa but non-linear movement (37.2 %). While SLC has been implemented for sperm separation in suboptimal and/or low concentrated sperm samples, this trial demonstrates that SLC is not efficient to separate different sperm populations in normospermic ram sperm samples containing high concentrations of spermatozoa

Comparing welfare indicators in dairy cattle under different loose housing systems (deep litter vs cubicle barns) using recycled manure solids for bedding

Aim of study: Dairy farmers in Southern Spain are continuously investing in the modernization of their facilities and frequently ask technicians about the type of housing they should choose for their farms. Although some studies have analysed the economic impact of different housing systems, there are no reports evaluating the impact of these systems on animal wellbeing. To remedy this deficit, a study was carried out to analyse animal welfare status in two types of loose housing conditions: deep litter (DL) and cubicle (CU) barns.Area of study: This study was conducted in Cordoba (Spain).Material and methods: A total of 1,597 cows from nineteen commercial dairy farms were involved in this study, of which twelve had CU barns and seven had DL barns. Welfare Quality assessment was used to evaluate animal wellbeing, inn order to compare both housing systems.Main results: The study found some weaknesses for feeding and health indicators of animal welfare in both types of housing systems. The overall welfare assessment based on feeding, housing and health indicators showed no differences between farms with DL or CU barns.Research highlights: A good welfare status could be reached under any type of housing system

Niveles químicos plasmáticos en vacas repetidoras tras I.A

Numerosos trabajos demuestran que durante el periodo de transición de la vaca aparecen desequilibrios metabólicos importantes que repercuten sobre la reproducción. Sin embargo, pocos hacen referencia al periodo temprano de gestación, donde es posible que, entre otras, las alteraciones o modificaciones en algunos constituyentes plasmáticos estén relacionadas con la repetición de celos. En dicho periodo existen grandes cambios hormonales, como los que suceden durante la formación del CL y subsecuente secreción de progesterona. Con el objetivo de ahondar en estos aspectos se llevó a cabo una experiencia en la que se determinaron las concentraciones de calcio, fósforo inorgánico, glucosa, magnesio, cobre y zinc presentes en el plasma sanguíneo de 42 vacas Holstein Freisian durante 32 días posteriores a la IA. Para determinar dichos parámetros metabólicos se empleó un fotómetro BioSystem BTS 310. Se realizaron análisis de varianza entre los animales agrupados según su estado reproductivo y según el número de IAs recibidas. Las concentraciones químicas determinadas en este estudio no sufrieron variaciones dependientes del estado reproductivo tras la IA, lo que indica que durante este periodo temprano se mantiene una homeostasis adecuada independientemente de los procesos de reconocimiento materno de la gestación que están en marcha o de variaciones hormonales que tienen lugar. Sin embargo, al comparar los resultados obtenidos entre las vacas repetidoras y las gestantes según el número de IAs recibidas se observan variaciones significativas en cuanto a los niveles de calcio, fósforo, glucosa, magnesio y zinc, lo que hace pensar en su posible participación en el denominado síndrome de la vaca repetidora

Factors influencing the success of an artificial insemination program in Florida goats

An artificial insemination (AI) program using cooled semen was evaluated over a 7-year period in Florida goats. The effect of the following variables on pregnancy rates was analysed: production system, year and season of AI, synchronisation treatment, bucks, AI technicians, semen deposition site, days in milk at AI, milk yield and parity. Animals were reared under field conditions on commercial farms in southern Spain. Semen was collected from nine bucks and cooled at 4ºC until use. A total of 3941 goats were synchronised using intravaginal progesterone sponges and inseminated 46.0 ± 0.5 h. after sponge removal. Pregnancy was diagnosed by ultrasonography 42-46 days after AI, obtaining an average pregnancy rate of 48.7%. Logistic regression showed that production system, AI year and season, bucks and semen deposition site had a significant effect (p<0.05) on pregnancy rate, while the other variables analysed were removed from the model. The final statistical model accounted for 59.7% of the cases analysed, suggesting that other factors not studied here may influence pregnancy rates in Florida goats

Evaluación ecográfica de la enfermedad quística ovárica en pequeños rumiantes

Se ha realizado un estudio de las características ecográficas de estructuras quísticas ováricas en 194 ovejas y 39 cabras de diferentes razas y procedentes de distintas zonas geográficas, con el objeto de valorar la ecografía transrectal con sondas de alta resolución (7,5 MHz) como criterio de selección de animales destinados a programas de superovulación y recogida de embriones. Los quistes foliculares tenían una incidencia de presentación del 4,1 p. 100 en las ovejas y del 5,1 p. 100 en las cabras, presentándose ecográficamente con una morfología esférica anecoica de 1,5±0,4 cm de diámetro, con paredes finas y generalmente bien definidas. Los quistes luteínicos tuvieron una presentación del 1 p. 100 en las ovejas y de¡ 2,6 p. 100 en las cabras. Ecográficamente se caracterizaban por presentar una cavidad central anecógena 1±0,2 cm, rodeada por un anillo ecogénico de 2-3 mm de espesor. No había diferencias significativas en la frecuencia de presentación de quistes ováricos entre ambas especies. Los resultados muestran que la ecografía transrectal con sondas de alta resolución permite la realizaciónde un diagnóstico eficazde la patología quística ovárica en los pequeños rumiantes, siendo recomendable esta metodología en la selección de hembras donantes y receptoras